ABSTRACT: Background It remains unknown whether individuals who regularly use preventive medications receive the same benefit from healthy lifestyle as those who do not use medications. We aimed to examine the associations of healthy lifestyle with mortality according to use of major preventive medications, including aspirin, antihypertensives, and lipid-lowering medications. Methods and Results Among 79 043 women in the Nurses' Health Study (1988-2014) and 39 544 men in the Health Professionals Follow-up Study (1986-2014), we defined a healthy lifestyle score based on body mass index, smoking, physical activity, diet, and alcohol intake. We estimated multivariable hazard ratios (HRs) and population-attributable risks of death from any cause, cardiovascular disease, cancer, and other causes in relation to healthy lifestyle according to medication use. We documented 35 195 deaths. A similar association of healthy lifestyle score with lower all-cause mortality was observed among medication users (HR, 0.82 per unit increment; 95% CI, 0.81-0.82) and nonusers (HR, 0.81; 95% CI, 0.79-0.83) (P interaction=0.54). The fraction of premature deaths that might be prevented by adherence to the 5 healthy lifestyle factors among medication users and nonusers was 38% (95% CI, 32%-42%) and 40% (95% CI, 29%-50%) for all-cause mortality, 37% (95% CI, 27%-46%) and 45% (95% CI, 18%-66%) for cardiovascular disease mortality, and 38% (95% CI, 28%-46%) and 33% (95% CI, 14%-49%) for cancer mortality, respectively. Conclusions Adherence to a healthy lifestyle confers substantial benefit for prevention of premature death among both regular users and nonusers of preventive medications. Adherence to a healthy lifestyle remains important even among individuals regularly using preventive medications.